Ifn αa

BMDC were generated as described (22 (link)). CD8⁺ T cells were isolated from spleen and lymph nodes of indicated mice through MACS-beads (Miltenyi Biotech). BMDC and T cell co-culture was performed as previously described (6 (link)). Briefly, we mixed DC with CD8⁺ T cells at the ratio of 1 DC : 10 T cells in round-bottom 96 wells (5 × 10⁴ T cells/well) in the presence of 0.1 µg/ml α-CD3. The conditions of the culture are indicated in the text. For RNA isolation and RT-PCR, T cells were harvested at day 3 of culture. For IFN-γ and IL-10 staining, T cells were harvested at day 4 of the culture and restimulated with PMA (100 ng/ml, Sigma) or inomycin (1 µg/ml, Sigma) in the presence of Golgi-Stop (1 µl/ml, BD Biosciences) for 4 hours. Intracellular cytokine staining (ICS) was performed as we previously reported (6 (link)). To measure T cell cytokine production by ELISA, day 4 cultured T cells were restimulated with plate bound α-CD3 (1 µg/ml) for overnight. Supernatants were then measured by ELISA as previously described (6 (link)). The concentrations of the cytokines used in the culture are as following: recombinant human IL-2 (300 U/ml); mouse IL-27 (Biolegend), 10 ng/ml and IFN-αA (R&D) 250 U/ml respectively.